In the detergent industry enzymes have for more than 30 years been implemented in washing formulations. Enzymes used in such formulations comprise proteases, lipases, amylases, cellulases, mannosidases as well as other enzymes or mixtures thereof. Commercially most important enzymes are proteases.
An increasing number of commercially used proteases is protein engineered variants of naturally occurring wild type proteases, e.g. DURAZYM® (Novozymes A/S), RELASE® (Novozymes A/S), MAXAPEM® (Gist-Brocades N.V.), PURAFECT® (Genencor International, Inc.).
Further, a number of protease variants is described in the art, such as in EP 130756 (GENENTECH) (corresponding to U.S. Re. Pat. No. 34,606 (GENENCOR)); EP 214435 (HENKEL); WO 87/04461 (AMGEN); WO 87105050 (GENEX); EP 260105 (GENENCOR); Thomas, Russell, and Fersht (1985) Nature 318 375-376; Thomas, Russell, and Fersht (1987) J. Mol. Biol. 193 803-813; Russel and Fersht Nature 328 496-500 (1987); WO 88/08028 (Genex); WO 88/08033 (Amgen); WO 95/27049 (SOLVAY S. A.); WO 95/30011 (PROCTER & GAMBLE COMPANY); WO 95/30010 (PROCTER & GAMBLE COMPANY); WO 95/29979 (PROCTER & GAMBLE COMPANY); U.S. Pat. No. 5,543,302 (SOLVAY S. A.); EP 251 446 (GENENCOR); WO 89/06279 (NOVOZYMES A/S); WO 91/00345 (NOVOZYMES A/S); EP 525 610 A1 (SOLVAY); WO 94/02618 (GIST-BROCADES N.V.).
Test method used for screening (AMSA) is described in WO 02/42740 (NOVOZYMES A/S).
WO 01/75087 (MAXYGEN, INC./NOVOZYMES A/S) describes subtilisin homologues that are improved for a variety of specific properties including thermal stability, activity at low temperature and alkaline stability.
WO 01/68821(NOVOZYMES A/S) describes subtilase enzymes, which are suitable for removal of egg stains from for example laundry and/or hard surfaces.
However, even though a number of useful proteases and protease variants have been described, there is still a need for further improvement of proteases or protease variants for a number of industrial uses.
In particular, the problem of removing egg stains from e.g. laundry or hard surfaces has been pronounced due to the fact that substances present in the egg white inhibit many serine proteases.
Therefore, an object of the present invention is to provide improved subtilase enzymes, which are suitable for removal of egg stains from for example laundry and/or hard surfaces.